Lung Tissue Culture Research Articles

Aspiration pneumonia in an American black bear

Information regarding American black bear (Ursus americanus) mortality caused by disease is limited. Pneumonia is a common respiratory disease that affects many species of wildlife and can result in death. In February 2011, we investigated the death of a yearling male black bear and determined cause of death to be aspiration pneumonia. We found sections of lung were diffusely congested and edematous. A bacterial culture of lung tissue revealed numerous colonies of Klebsiella spp., Alcaligenes faecalis, Aeromonas spp., Corynebacterium spp., and Streptococcus spp. The mixed bacterial colonization of the lungs associated with minimal inflammation is consistent with terminal aspiration of gastrointestinal contents resulting in aspiration pneumonia. Documentation of aspiration pneumonia in black bears can be useful for future researchers interested in the effects of disease on bears.

The role of postmortem studies in pneumonia etiology research.

The diagnosis of etiology in severe pneumonia remains a challenging area. Postmortem lung tissue potentially increases the sensitivity of investigations for identification of causative pathogens in fatal cases of pneumonia and can confirm antemortem microbiological diagnoses. Tissue sampling allows assessment of histological patterns of disease and ancillary immunohistochemical or molecular diagnostic techniques. It may also enhance the recognition of noninfectious conditions that clinically simulate acute pneumonia. Biobanking of lung tissue or postmortem culture isolates offers opportunities for new pathogen discovery and research into host-pathogen interactions. The Pneumonia Etiology Research for Child Health study proposes a percutaneous needle biopsy approach to obtain postmortem samples, rather than a full open autopsy. This has the advantage of greater acceptability to relatives, but risks greater sampling error. Both approaches may be susceptible to microbiological contamination or pathogen degradation. However, previous autopsy studies have confirmed the value of histological examination in revealing unsuspected pathogens and influencing clinical guidelines for the diagnosis and treatment of future pneumonia cases.

Sec62 Bridges the Gap from 3q Amplification to Molecular Cell Biology in Non–Small Cell Lung Cancer

The molecular carcinogenesis of lung cancer has yet to be clearly elucidated. We investigated the possible oncogenic function of SEC62 in lung cancer, which was predicted based on our previous findings that lung and thyroid cancer tissue samples exhibited increased Sec62 protein levels. The SEC62 gene locus is at 3q26.2, and 3q amplification is reportedly the most common genomic alteration in non-small cell lung cancer. We analyzed SEC62 mRNA and protein levels in tissue samples from lung cancer patients by real-time quantitative PCR, Western blot, and IHC and found significantly increased SEC62 mRNA and protein levels in tumors compared with tumor-free tissue samples from the same patients. Correlation analyses revealed significantly higher Sec62 levels in tumors with lymph node metastases compared with nonmetastatic tumors, as well as in poorly compared with moderately differentiated tumors. On the basis of these promising results, we examined the role of Sec62 in cancer cell biology in vitro. Cell migration assays with lung and thyroid cancer cells showed distinct stimulation of migration in SEC62-overexpressing cells and inhibition of migration in Sec62-depleted cells. Moreover, we found that SEC62 silencing sensitized the cells to thapsigargin-induced endoplasmic reticulum stress. Thus, our results indicate that SEC62 represents a potential candidate oncogene in the amplified 3q region in cases of non-small cell lung cancer and harbors various functions in cancer cell biology.

Bioreactor for the Long-Term Culture of Lung Tissue

In this article we describe the design and validation of a bioreactor for the in vitro culture of whole rodent lung tissue. Many current systems only enable large segments of lung tissue to be studied ex vivo for up to a few hours in the laboratory. This limitation restricts the study of pulmonary biology in controlled laboratory settings, and also impacts the ability to reliably culture engineered lung tissues in the laboratory. Therefore, we designed, built, and validated a bioreactor intended to provide sufficient nutrient supply and mechanical stimulation to support cell survival and differentiation in cultured lung tissue. We also studied the effects of perfusion and ventilation on pulmonary cell survival and maintenance of cell differentiation state. The final bioreactor design described herein is capable of supporting the culture of whole native lung tissue for up to 1 week in the laboratory, and offers promise in the study of pulmonary biology and the development of engineered lung tissues in the laboratory.

Mycobacterium mageritensePulmonary Disease in Patient with Compromised Immune System

<i>Mycobacterium mageritense</i>Pulmonary Disease in Patient with Compromised Immune System

Oxidative modification of albumin in the parenchymal lung tissue of current smokers with chronic obstructive pulmonary disease

BackgroundThere is accumulating evidence that oxidative stress plays an important role in the pathophysiology of chronic obstructive pulmonary disease (COPD). One current hypothesis is that the increased oxidant burden in these patients is not adequately counterbalanced by the lung antioxidant systems.ObjectiveTo determine the levels of oxidised human serum albumin (HSA) in COPD lung explants and the effect of oxidation on HSA degradation using an ex vivo lung explant model.MethodsParenchymal lung tissue was obtained from 38 patients (15F/23M) undergoing lung resection and stratified by smoking history and disease using the GOLD guidelines and the lower limit of normal for FEV1/FVC ratio. Lung tissue was homogenised and analysed by ELISA for total levels of HSA and carbonylated HSA. To determine oxidised HSA degradation lung tissue explants were incubated with either 200 μg/ml HSA or oxidised HSA and supernatants collected at 1, 2, 4, 6, and 24 h and analysed for HSA using ELISA and immunoblot.ResultsWhen stratified by disease, lung tissue from GOLD II (median = 38.2 μg/ml) and GOLD I (median = 48.4 μg/ml) patients had lower levels of HSA compared to patients with normal lung function (median = 71.9 μg/ml, P < 0.05). In addition the number of carbonyl residues, which is a measure of oxidation was elevated in GOLD I and II tissue compared to individuals with normal lung function (P < 0.05). When analysing smoking status current smokers had lower levels of HSA (median = 43.3 μg/ml, P < 0.05) compared to ex smokers (median = 71.9 μg/ml) and non-smokers (median = 71.2 μg/ml) and significantly greater number of carbonyl residues per HSA molecule (P < 0.05). When incubated with either HSA or oxidised HSA lung tissue explants rapidly degraded the oxidised HSA but not unmodified HSA (P < 0.05).ConclusionWe report on a reliable methodology for measuring levels of oxidised HSA in human lung tissue and cell culture supernatant. We propose that differences in the levels of oxidised HSA within lung tissue from COPD patients and current smokers provides further evidence for an oxidant/antioxidant imbalance and has important biological implications for the disease.

Hepatocyte Growth Factor Inhibits Apoptosis by the Profibrotic Factor Angiotensin II via Extracellular Signal-regulated Kinase 1/2 in Endothelial Cells and Tissue Explants

Hepatocyte growth factor (HGF), an endogenous tissue repair factor, attenuates apoptosis in many primary cell types, but the mechanism is not completely understood. Our laboratory demonstrated that angiotensin (Ang) II activates the intrinsic apoptotic pathway in primary endothelial cells (ECs) via reduction of the antiapoptotic protein Bcl-x(L). Ang II decreased Bcl-x(L) mRNA half-life by reducing its binding to nucleolin, a protein that normally binds a 3' AU-rich region and stabilizes Bcl-x(L) mRNA. We hypothesized HGF may block apoptosis induced by Ang II. We used primary EC and ex vivo cultures of rat lung tissue to investigate HGF inhibition of Ang II-induced apoptosis. Our data indicated HGF abrogated Ang II-induced apoptosis by inhibiting cytochrome c release, caspase-3 activation, and DNA fragmentation. RNA-immunoprecipitation experiments demonstrated that HGF stabilized Bcl-x(L) mRNA by increasing nucleolin binding to the 3'-untranslated region that was associated with cytoplasmic localization of nucleolin. Cytoplasmic localization of nucleolin and Bcl-x(L) mRNA stabilization required HGF activation of extracellular signal-regulated kinase (ERK)1/2, but not phosphatidylinositol 3-kinase. HGF also blocked Ang II-induced caspase-3 activation and lactate dehydrogenase release in tissue explants in an ERK-dependent manner.

Aspergillus niger: an unusual cause of invasive pulmonary aspergillosis

Infections due to Aspergillus species cause significant morbidity and mortality. Most are attributed to Aspergillus fumigatus, followed by Aspergillus flavus and Aspergillus terreus. Aspergillus niger is a mould that is rarely reported as a cause of pneumonia. A 72-year-old female with chronic obstructive pulmonary disease and temporal arteritis being treated with steroids long term presented with haemoptysis and pleuritic chest pain. Chest radiography revealed areas of heterogeneous consolidation with cavitation in the right upper lobe of the lung. Induced bacterial sputum cultures, and acid-fast smears and cultures were negative. Fungal sputum cultures grew A. niger. The patient clinically improved on a combination therapy of empiric antibacterials and voriconazole, followed by voriconazole monotherapy. After 4 weeks of voriconazole therapy, however, repeat chest computed tomography scanning showed a significant progression of the infection and near-complete necrosis of the right upper lobe of the lung. Serum voriconazole levels were low–normal (1.0 μg ml−1, normal range for the assay 0.5–6.0 μg ml−1). A. niger was again recovered from bronchoalveolar lavage specimens. A right upper lobectomy was performed, and lung tissue cultures grew A. niger. Furthermore, the lung histopathology showed acute and organizing pneumonia, fungal hyphae and oxalate crystallosis, confirming the diagnosis of invasive A. niger infection. A. niger, unlike A. fumigatus and A. flavus, is less commonly considered a cause of invasive aspergillosis (IA). The finding of calcium oxalate crystals in histopathology specimens is classic for A. niger infection and can be helpful in making a diagnosis even in the absence of conidia. Therapeutic drug monitoring may be useful in optimizing the treatment of IA given the wide variations in the oral bioavailability of voriconazole.

Viral respiratory tract infections and asthma: The course ahead

Inquiries into the relationships between viral respiratory tract illnesses and the inception and exacerbation of asthma are being facilitated by recent advances in research approaches and technology. In this article we identify important knowledge gaps and future research questions, and we discuss how new investigational tools, including improved respiratory tract virus detection techniques, will permit current and future researchers to define these relationships and the host, virus, developmental, and environmental mechanisms that regulate them. A better understanding of these processes should facilitate the development of improved strategies for the prevention and treatment of virus-induced wheezing illnesses and asthma exacerbations and, possibly, the ultimate goal of discovering effective approaches for the primary prevention of asthma.

The TGF-beta-Pseudoreceptor BAMBI is strongly expressed in COPD lungs and regulated by nontypeable Haemophilus influenzae

BackgroundNontypeable Haemophilus influenzae (NTHI) may play a role as an infectious trigger in the pathogenesis of chronic obstructive pulmonary disease (COPD). Few data are available regarding the influence of acute and persistent infection on tissue remodelling and repair factors such as transforming growth factor (TGF)-β.MethodsNTHI infection in lung tissues obtained from COPD patients and controls was studied in vivo and using an in vitro model. Infection experiments were performed with two different clinical isolates. Detection of NTHI was done using in situ hybridization (ISH) in unstimulated and in in vitro infected lung tissue. For characterization of TGF-β signaling molecules a transcriptome array was performed. Expression of the TGF-pseudoreceptor BMP and Activin Membrane-bound Inhibitor (BAMBI) was analyzed using immunohistochemistry (IHC), ISH and PCR. CXC chemokine ligand (CXCL)-8, tumor necrosis factor (TNF)-α and TGF-β expression were evaluated in lung tissue and cell culture using ELISA.ResultsIn 38% of COPD patients infection with NTHI was detected in vivo in contrast to 0% of controls (p < 0.05). Transcriptome arrays showed no significant changes of TGF-β receptors 1 and 2 and Smad-3 expression, whereas a strong expression of BAMBI with upregulation after in vitro infection of COPD lung tissue was demonstrated. BAMBI was expressed ubiquitously on alveolar macrophages (AM) and to a lesser degree on alveolar epithelial cells (AEC). Measurement of cytokine concentrations in lung tissue supernatants revealed a decreased expression of TGF-β (p < 0.05) in combination with a strong proinflammatory response (p < 0.01).ConclusionsWe show for the first time the expression of the TGF pseudoreceptor BAMBI in the human lung, which is upregulated in response to NTHI infection in COPD lung tissue in vivo and in vitro. The combination of NTHI-mediated induction of proinflammatory cytokines and inhibition of TGF-β expression may influence inflammation induced tissue remodeling.

Tissue engineering and biotechnology in general thoracic surgery

Public interest in the recent progress of tissue engineering, a special line of biotechnology, makes the current review on thoracic surgery highly relevant. In this article, techniques, materials and cellular processes are discussed alongside their potential applications in tissue repair. Different applications of tissue engineering in tracheo-bronchial replacement, lung tissue cultures and chest-wall reconstruction are also summarised in the article. Potential tissue engineering-based solutions for destructive, chronic lung-injury-related conditions and replacement of tubular structures in the central airways are also examined.

Enterococcus faecium lung abscess: one case report and literature review

to study the diagnosis and treatment of enterococcus faecium lung abscess. a retrospective analysis of one case of Enterococcus faecium lung abscess and literature review was conducted. this patient suffered from cough and sputum over 6 months and complicated with hemoptysis over 3 months. Pulmonary embolism and lung cancer were suspected initially. After 2 times of CT-guided percutaneous transthoracic needle aspiration biopsy the diagnosis of pneumonia was made in other hospitals. However, the consolidation in the lung progressed and cavity appeared although antibiotic therapy was conducted. After admission to our hospital, CT-guided percutaneous transthoracic needle aspiration biopsy was made and the lung tissue was sent for bacterial culture. Enterococcus faecium was cultured and it was susceptible to vancomycin, teicoplanin and linezolid. The disease improved significantly after treatment with these 3 antibiotics in turn. In addition, 13 cases of enterococcus pneumonia or lung abscess were reviewed, including 3 cases of enterococcus faecium lung abscess. enterococcus faecium is rarely a pathogen for lung abscess. The diagnosis of enterococcus faecium lung abscess could be confirmed by lung biopsy and bacterial culture of lung tissue which could also provide the susceptibility of antibiotics and guide the antibiotic therapy.

A rare presentation of aspergillus infection as empyema thoracis

A 57-year-old diabetic and hypertensive man presented with a short history of fever, dry cough and right side chest pain. A chest radiograph showed right pleural based homogenous shadow in middle and lower zones with obliteration of right costo-phrenic angle suggestive of right side effusion. Aspiration of pleural fluid revealed frank pus for which inter-costal tube drainage was performed. Due to persistence of empyema, the patient was subjected to thoracoscopy. Thoracoscopy showed multiloculated empyema. Thoracoscopic pleural biopsy and fluid showed septate fungal hyphae. Thoracotomy and parietal pleurectomy, with resection of part of right lower lobe, was carried out. Pleural fluid, pleural and lung tissue culture grew Aspergillus fumigatus. The patient showed good recovery with voriconazole after thoracotomy.

Chorioamnionitis and Neonatal Sepsis from Community-associated MRSA

Chorioamnionitis and Neonatal Sepsis from Community-associated MRSA

The karyotype of the grey seal (Halichoerus grypus).

The somatic chromosomes of the grey seal, Halichoerus grypus Fabr., and their autoradiographic pattern were studied in cultures of lung tissue. An idiogram was made based on measurements of 6 male and 14 female cells. The chromosome number of the grey seal is 2n = 32. Some comments were made on the karyological interrelationships among the species so far studied of the family Phocidae.

DAS181 inhibits H5N1 influenza virus infection of human lung tissues.

DAS181 is a novel candidate therapeutic agent against influenza virus which functions via the mechanism of removing the virus receptor, sialic acid (Sia), from the adjacent glycan structures. DAS181 and its analogues have previously been shown to be potently active against multiple strains of seasonal and avian influenza virus strains in several experimental models, including cell lines, mice, and ferrets. Here we demonstrate that DAS181 treatment leads to desialylation of both alpha2-6-linked and alpha2-3-linked Sia in ex vivo human lung tissue culture and primary pneumocytes. DAS181 treatment also effectively protects human lung tissue and pneumocytes against the highly pathogenic avian influenza virus H5N1 (A/Vietnam/3046/2004). Two doses of DAS181 treatment given 12 h apart were sufficient to block H5N1 infection in the ex vivo lung tissue culture. These findings support the potential value of DAS181 as a broad-spectrum therapeutic agent against influenza viruses, especially H5N1.

Characterization of late tuberculosis infection in Sigmodon hispidus

We previously described primary tuberculosis in Sigmodon hispidus cotton rats up to 6 months following a pulmonary challenge. At that time, we observed fewer animals demonstrating disease as time from exposure progressed. We hypothesized that some cotton rats may control a primary infection to latency in a similar fashion to humans. The current experiment was designed to examine the natural progression of disease in S. hispidus at a later timepoint following a respiratory challenge with Mycobacterium tuberculosis (Mtb). An additional objective was to test whether cotton rats may become latently infected, and to determine whether latent disease might be activated by cyclophosphamide induced immune suppression. Thirty-four percent of the inoculated cotton rats died prior to 9 months following the challenge. However, 50% of immunocompetent animals surviving past 9 months demonstrated positive lung tissue cultures for Mtb without histologic evidence of disease. None of the immunosuppressed animals demonstrated this pattern. These findings are consistent with the development of latent tuberculosis infection in some cotton rats. Furthermore, it appears reactivation of disease occurs with cyclophosphamide induced immunosuppression. Cotton rats may serve as a model for latent as well as active tuberculosis infection.

Development of a nested PCR assay for the detection of Fusarium solani DNA and its evaluation in the diagnosis of invasive fusariosis using an experimental mouse model

Fusarium infections are increasingly being encountered in immunocompromised patients. Fusarium solani accounts for nearly half of these infections. A specific nested PCR (nPCR) assay has been developed by using DNA isolated from several Fusarium species and other common fungi. Furthermore, DNA samples isolated from bronchoalveolar lavage (BAL) and serum samples from mice infected intravenously with F. solani conidia and sacrificed on every third day post infection were used for the evaluation of the established nPCR protocol. The lung homogenate, BAL and blood from infected animals were also cultured. The nPCR assay was specific for F. solani and detected 450 fg of DNA corresponding roughly to 11 F. solani cells. Cultures of lung homogenate of infected animals up to day 16 yielded F. solani with decreasing fungal load and were negative thereafter. The nPCR positivity in BAL was 100% concordant with lung tissue culture results. Although detection of F. solani DNA in serum was less sensitive than in BAL, it remained positive for longer duration. Our data from an experimental mouse model show that detection of DNA in BAL and to a lesser extent in serum by nPCR offers a sensitive and specific diagnostic approach to invasive F. solani infection.

Recurrent Bilateral Pneumothoraces: A Rare Complication of Miliary Tuberculosis

Pneumothorax as a complication of adult cavitary pulmonary tuberculosis is well known and not at all rare, but its occurrence as a complication of miliary tuberculosis is extremely rare. We report a 22-year-old woman who had nonproductive cough and fever for 3 days. Chest radiography showed diffuse, symmetrical miliary nodulation throughout both lung fields. The patient was treated for a presumed diagnosis of miliary tuberculosis with standard antituberculous regimen. Bilateral pneumothorax occurred simultaneously during hospitalization and chest tube thoracostomy was performed. Three days later, recurrent right pneumothorax developed. Video-assisted thoracoscopic surgery (VATS) lung biopsy of the right lung was performed and pathology showed granulomatous interstitial pneumonia with acid-fast positive bacilli. Lung tissue culture was positive for Mycobacterium tuberculosis. In the following 2 months, bilateral pneumothorax recurred twice and chemical pleurodesis with minocycline was performed on both sides, but air leakage persisted. VATS pleurodesis was performed on both sides successfully without recurrence of pneumothorax on either side. Our experience highlights the fact that pneumothorax should be suspected in an adult with miliary tuberculosis who suddenly develops acute respiratory distress. Recurrent pneumothorax can be managed, apart from medical therapy of miliary tuberculosis, with surgical intervention.

Sarcoidosis Presenting With Cystic Lung Lesions and Autoimmune Hemolytic Anemia

Introduction Hemolytic anemia is an unusual finding in sarcoidosis. We report an intriguing case of hemolytic anemia in cystic sarcoidosis. Case Presentation A 34-year-old African American male with bronchial asthma presented with wheezing, exertional dyspnea, cough, fever, night sweats, weight loss, and fatigue. Physical examination revealed pallor, palpable peripheral adenopathy, and expiratory rhonchi. Initial laboratory findings were remarkable for leukopenia, anemia, reticulocytosis, elevated lactate dehydrogenase, and abnormal liver function test. Serum haptoglobin was low. Sickling test was negative. Direct Coombs test was positive, and cold agglutinins were demonstrated. Chest radiograph revealed cystic lesions in both upper lung fields, and chest computed tomographic scan confirmed these lesions. Also seen were mediastinal adenopathy and hepatosplenomegaly. Tuberculin skin test was negative in 48 hours. Three sputum smears for acid-fast bacilli were negative. Serum angiotensin converting enzyme level was elevated. He underwent diagnostic bronchoscopy with carinal and transbronchial lung biopsies revealing non-necrotizing granulomatous inflammation consistent with sarcoidosis. Acid-fast bacilli and silver stains were negative. Respiratory and lung tissue cultures were negative for fungi and Mycobacteria. Diagnosis of sarcoidosis with autoimmune hemolytic anemia was established and systemic corticosteroid was instituted with resolution of hemolysis and modest roentgenographic regression of the cystic lung lesions. Conclusions Some cases of cystic/bullous sarcoidosis had been reported and the occurrence of autoimmune disease with sarcoidosis well known. To the best of our knowledge, this is the first reported case of sarcoidosis presenting with cystic lung lesions and autoimmune hemolytic anemia in a single patient.